1. APPLICATION OF
CHROMATOGRAPHY
Presented by:
Mr. Shankar Yelmame
S.Y.M.pharm

2. CONTENTS
 Types & application of Chromatography
1) HPLC
2) LC-MS
3) HPTLC
4) GC
5) GC-MS
6) Supercritical fluid chromatography
7) UPLC

3. High Performance Liquid
Chromatography
1) Natural products
- Separation of components in plant extract
- nucleic acids, urine, serum, carbohydrates, lipids,
amino acids, bile salts etc.
2) Separation of oligopeptides & proteins
3) Stability studies
-analysis of degradation products
4) Bio-assay & its complementation
- antibiotics & peptide hormones

4. 5) Cosmetic industries
- Lipstics

5. LC-MS
Liquid chromatography–mass spectrometry
1) Most frequently used technique in the field of bioanalysis.
2) Components of a complex mixture must be detected and identified.
3) LC-MS is frequently used in drug development at many different
stages including
- peptide mapping,
- glycoprotein mapping,
- natural products de-replication,
- in vivo drug screening,
- impurity identification,
-quantitative bio-analysis and quality control

6. HPTLC
High performance thin layer chromatography
1) Used in analyzing the purity and efficacy of many
pharmaceutical preparations and dosage forms.
2) It is also an ideal screening tool for adulterations.
3) HPTLC is used for purity control of chemicals,
pesticides, steroids, and water analysis.
4) Phytochemical analysis, biomedical analysis, herbal
drug quantification.

7. GAS CHROMATOGRAPHY
1) Qualitative analysis :
a) By comparing the retention times/volumes of the
unknown with the standard.
b) By comparing the individual components.
2) Quantitative analysis :
- depends upon the fact that the area unde a single
component peak is proportional to quantity of the detected
component.

8. 3) Miscellaneous :
a) Detection of steroids
b) Analysis of foods
c) Separation & identification of lipids, proteins,
carbohydrates, vitamins, pesticides .
d) Analysis of dairy products
e) Separation & identification of volatile materials like
plastics, natural & synthetic polymers.

9. GC-MS
Gas chromatography–mass spectrometry
1) Sports anti-doping analysis
-GC-MS is the main tool used in sports anti-doping laboratories
to test athletes' urine samples for prohibited performance-
enhancing drugs.
e.g. anabolic steroids
2) Food, beverage and perfume analysis.
3) Law enforcement
-GC-MS is increasingly used for detection of illegal narcotics.

10. 4) Separation & identification of degradation products of
organic & organometallics making the elucidation of their
stuctures.
5) Diagnosis of inborn errors of metabolism in New borns.

11. Supercritical Fluid
Chromatography
 SFC is used in industry primarily for separation of chiral
molecules,
 SFC is now commonly used for achiral separations and
purifications in the pharmaceutical industry.

12. UPLC
Ultra Performance Liquid Chromatography
1. LOW SAMPLE CONSUMPTION
2. TIME SAVING
3. DRUG DISCOVERY
4. ANALYSIS OF ENVIRONMENTAL RESIDUES
5. SEPARTION OF AMINO ACID

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